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Molecular markers for detecting carbostyril-resistant gene of bovine-derived Escherichia coli and application thereof

A technology of Escherichia coli and molecular marker, applied in the field of molecular biology, can solve problems such as economic loss of animal husbandry, and achieve the effects of strong sensitivity, good stability and high specificity

Inactive Publication Date: 2017-10-03
SUZHOU JONATHAN NEW MATERIALS TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since Escherichia coli is often concurrent or secondary to other viral or bacterial diseases, antibacterial drugs are often used as feed additives in livestock and poultry farming, which often cause serious economic losses to the livestock industry

Method used

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  • Molecular markers for detecting carbostyril-resistant gene of bovine-derived Escherichia coli and application thereof

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Experimental program
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Effect test

Embodiment 1

[0026] A molecular marker for detecting the quinolone drug-resistant gene of bovine Escherichia coli, the molecular marker and its sequence are: P1, SEQ ID NO.1-2; P2, SEQ ID NO.3-4; P3, SEQ ID NO.5~6.

[0027] The 5' ends of the upstream primers of the molecular markers P1, P2 and P3 were modified by amination and 10 bases T were added.

[0028] The application of the molecular marker in the preparation of a kit for detecting bovine Escherichia coli quinolone drug resistance gene.

[0029] The components of the kit include: molecular markers, dNTP, LATaq, PCR reaction buffer, MgCl 2 , double distilled water.

[0030] The steps of the kit detecting bovine Escherichia coli quinolone drug resistance gene include:

[0031] (1) Sampling: Select purified bovine Escherichia coli, streak inoculate the culture medium to form a single colony, add 10 μL of 60% glycerol to a sterile EP tube, pick a single colony with an inoculation needle and wash it several times in glycerin , store...

Embodiment 2

[0041] A molecular marker for detecting the quinolone drug-resistant gene of bovine Escherichia coli, the molecular marker and its sequence are: P1, SEQ ID NO.1-2; P2, SEQ ID NO.3-4; P3, SEQ ID NO.5~6.

[0042] The 5' ends of the upstream primers of the molecular markers P1, P2 and P3 were modified by amination and 10 bases T were added.

[0043] The application of the molecular marker in the preparation of a kit for detecting bovine Escherichia coli quinolone drug resistance gene.

[0044] The components of the kit include: molecular markers, dNTP, LATaq, PCR reaction buffer, MgCl 2 , double distilled water.

[0045] The steps of the kit detecting bovine Escherichia coli quinolone drug resistance gene include:

[0046] (1) Sampling: Select purified bovine Escherichia coli, inoculate the culture medium by streaking to form a single colony, add 30 μL of 60% glycerol to a sterile EP tube, pick a single colony with an inoculation needle and wash it several times in glycerin ,...

Embodiment 3

[0056] A molecular marker for detecting the quinolone resistance gene of bovine Escherichia coli, the molecular marker and its sequence are: P1, SEQ ID NO.1-2; P2, SEQ ID NO.3-4; P3, SEQ ID NO.5~6.

[0057] The 5' ends of the upstream primers of the molecular markers P1, P2 and P3 were modified by amination and 10 bases T were added.

[0058] The application of the molecular marker in the preparation of a kit for detecting bovine Escherichia coli quinolone drug resistance gene.

[0059] The components of the kit include: molecular markers, dNTP, LATaq, PCR reaction buffer, MgCl 2 , double distilled water.

[0060] The steps of the kit detecting bovine Escherichia coli quinolone drug resistance gene include:

[0061] (1) Sampling: Select purified bovine Escherichia coli, inoculate the culture medium by streaking to form a single colony, add 60 μL of 60% glycerol to a sterile EP tube, pick a single colony with an inoculation needle and wash it several times in glycerin , sto...

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Abstract

The invention discloses molecular markers for detecting a carbostyril-resistant gene of bovine-derived Escherichia coli and application thereof. The molecular markers have sequences of P1, SEQ ID NO. 1 to 2, P2, SEQ ID NO. 3 to 4 and P3, SEQ ID NO. 5 to 6. The molecular markers can detect production of a drug-resistant mutant strain in real time in bacterial infection treatment, can evaluate antibiotic potencies and a drug-resistant mutation prevention capability of the antibacterial agent and have the advantages of high specificity, high sensitivity and good stability.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to a method for detecting microbial drug resistance genes by means of molecular biology, in particular to a molecular marker for detecting bovine Escherichia coli quinolone drug resistance genes and an application thereof. Background technique [0002] In 1922, bacteriologists discovered a "lysozyme" from nasal mucus cultures, which is the prototype of penicillin. In 1927, Fleming officially published an article stating the discovery of penicillin. But it was not until 1939 that the Australian Ferroly and the German Chain En invented the penicillin purification technology, and two years later penicillin began to be officially used clinically. Since then, the treatment of bacterial diseases has entered a new era. In the following decades, the family of antibacterial drugs continued to grow, and new antibacterial drugs were continuously used in the treatment of infectious diseases, br...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/10C12N15/11C12R1/19
CPCC12Q1/689C12Q2600/106
Inventor 吕少波李苏杨李卓才
Owner SUZHOU JONATHAN NEW MATERIALS TECH
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